Interferon-Gamma-Inducible Protein 16 Inhibits Hepatocellular Carcinoma via Interferon Regulatory Factor 3 on Chemosensitivity.

BACKGROUND AND AIM: Previous reports have suggested IFI16 as a tumor suppressor in hepatocellular carcinoma (HC). Nonetheless, the biological significance of IFI16 and its mechanism concerning resistance to cisplatin (DDP) in HC requires further exploration. METHODS: Samples of tumor and corresponding para-carcinoma tissues were acquired from patients with HC. Furthermore, DDP-resistant cell lines of HC, specifically HCC, Huh7 and Hepatoblastoma, HepG3, were generated by gradually increasing the concentration of DDP. Cell apoptosis and DNA damage were evaluated by utilizing flow cytometry assay and TUNEL staining. The interaction between IFI16 and interferon regulatory factor 3 (IRF3) proteins were analyzed using Co-Immunoprecipitation (Co-IP) assay. In vivo assays were conducted by establishing HC subcutaneous xenograft tumor models. RESULTS: The study found a reduction in IFI16 expression in both HC tissues and DDP-resistant HC cell lines. The binding of IFI16 to IRF3 regulated DNA damage-associated markers in vitro. Overexpression of IFI16 heightened the susceptibility of DDP-induced apoptosis and DNA damage, which was counteracted by IRF3 knockdown, while strengthened by IRF3 overexpression. Moreover, overexpression of IFI16 diminished in vivo DDP-resistant HC tumorigenicity. CONCLUSION: In summary, our findings suggest that IFI16 serves as a tumor suppressor in HC by promoting DNA damage via its interaction with IRF3, thereby reversing DDP resistance.

Association of hysterectomy with nonalcoholic fatty liver disease among US women.

BACKGROUND: A postmenopausal rise in the rates of nonalcoholic fatty liver disease (NAFLD) has been reported in women. This study thus sought to further probe the association of hysterectomy with NAFLD. METHODS: The data utilized in this investigation were attained from the 2017-March 2020 cycle of the National Health and Nutrition Examination Survey (NHANES), reflecting a strategic utilization of comprehensive health and nutrition information in the US population, to conduct a cross-sectional examination of the relationship between self-reported hysterectomy and NAFLD. Subjects included in this study were women aged 20 years or older. The multivariable logistic regression methodologies were utilized to determine the pertinent odds ratios (ORs) and their associated 95% confidence intervals (CIs). RESULTS: Of the 2,868 subjects enrolled in this study (mean age: 51.3 years, 95%CI: 50.0-52.6 years), 22.1% (95%CI: 19.7-24.7%) reported having undergone a hysterectomy, while 31.1% (95%CI: 28.1-34.1%) exhibited elastographic evidence of NAFLD, and 3.8% (95%CI: 2.6-5.6%) exhibited clinically significant fibrosis (CSF). Relative to women with no history of hysterectomy, those that had undergone hysterectomy exhibited a higher odd of NAFLD (OR:1.66, 95%CI: 1.24-2.21) in a multivariable model fully adjusted for age, ethnicity, body mass index, female hormone use, oophorectomy, diabetes, hyperlipidemia, and smoking status. Subgroup analyses revealed a stronger association among women who were not obese (OR:2.23, 95%CI:1.61-3.11), women who were not affected by diabetes (OR:1.76, 95%CI: 1.25-2.46), and without hyperlipidemia (OR: 1.87, 95%CI: 1.10-3.16). No significant association of hysterectomy with NAFLD encompassing CSF was identified. CONCLUSIONS: The results of the present nationally representative analysis suggested an association between hysterectomy and increased NAFLD prevalence among US women. Knowledge of this relationship may better aid clinical efforts to screen for and manage NAFLD.

Multifunctional Metasurfaces Enabled by Multifold Geometric Phase Interference.

Geometric phase is frequently used in artificially designed metasurfaces; it is typically used only once in reported works, leading to conjugate responses of two spins. Supercells containing multiple nanoantennas can break this limitation by introducing more degrees of freedom to generate new modulation capabilities. Here, we provide a method for constructing supercells for geometric phases using triple rotations, each of which achieves a specific modulation function. The physical meaning of each rotation is revealed by stepwise superposition. Based on this idea, spin-selective holography, nanoprinting, and their hybrid displays are demonstrated. As a typical application, we have designed a metalens that enables spin-selective transmission, allowing for high-quality imaging with only one spin state, which can serve as a plug-and-play chiral detection device. Finally, we analyzed how the size of supercells and the phase distribution inside it can affect the higher order diffraction, which may help in designing supercells for different scenarios.

Characterization of Stromatolite Organic Sedimentary Structure Based on Spectral Image Fusion.

This paper evaluates the potential application of Raman baselines in characterizing organic deposition. Taking the layered sediments (Stromatolite) formed by the growth of early life on the Earth as the research object, Raman spectroscopy is an essential means to detect deep-space extraterrestrial life. Fluorescence is the main factor that interferes with Raman spectroscopy detection, which will cause the enhancement of the Raman baseline and annihilate Raman information. The paper aims to evaluate fluorescence contained in the Raman baseline and characterize organic sedimentary structure using the Raman baseline. This study achieves spectral image fusion combined with mapping technology to obtain high spatial and spectral resolution fusion images. To clarify that the fluorescence of organic matter deposition is the main factor causing Raman baseline enhancement, 5041 Raman spectra were obtained in the scanning area of 710 µm × 710 µm, and the correlation mechanism between the gray level of the light-dark layer of the detection point and the Raman baseline was compared. The spatial distribution of carbonate minerals and organic precipitations was detected by combining mapping technology. In addition, based on the BI-IHS algorithm, the spectral image fusion of Raman fluorescence mapping and reflection micrograph, polarization micrograph, and orthogonal polarization micrograph are realized, respectively. A fusion image with high spectral resolution and high spatial resolution is obtained. The results show that the Raman baseline can be used as helpful information to characterize stromatolite organic sedimentary structure.

Elastic Particle Swarm Optimization for MarSCoDe Spectral Calibration on Tianwen-1 Mars Rover.

China's Tianwen-1 Mars rover carries a laser-induced breakdown spectroscopy (LIBS) payload named MarSCoDe to analyze the mineral and rock composition on Mars. MarSCoDe is expected to experience a wide working temperature range of about 100 °C, which will lead to a spectral shift of up to ∼40 pixels (∼8.13 nm). Even worse, drastic changes in temperature and environment may cause a loss or increase of some spectral lines of an on-board calibration Ti target. An elastic particle swarm optimization (PSO) approach is proposed to fulfill the on-board spectral calibration of MarSCoDe under this harsh condition. Through establishing a standard wavelength set (SWS) and an individual particle wavelength set (PWS), and further elastically selecting a part of PWS to compare with SWS, the problem of spectral shift and number mismatch can be solved gradually with the evolution of the particle swarm. Some tests of standard lamps and Ti with MarSCoDe, placed in a Mars simulation environment chamber (MSEC) in a temperature range of 70 °C, were completed. Compared with the standard spectrum of the Ti target (obtained at 20 °C), the spectral shifts of the first, second, and third channels are approximately 0.33 nm (5 pixels), 0.85 nm (6.4 pixels), and 8.09 nm (39.8 pixels), respectively, at -40 °C before correction; after PSO correction, the spectral shifts are greatly reduced to up to 0.015 nm, and specially for the 626.28 nm line, the spectral shift is reduced from 8.09 nm to about 0 nm. Experimental results demonstrate that the PSO-based approach can not only correct the on-board spectral shift but also solve the number mismatch of spectral lines of MarSCoDe in the harsh working environment of Mars. Further, it can be extended to the on-board calibration of other spectral payloads for deep space exploration.

ASK1 Enhances Angiotensin II-Induced Liver Fibrosis In Vitro by Mediating Endoplasmic Reticulum Stress-Dependent Exosomes.

BACKGROUND: Apoptosis signal-regulating kinase 1 (ASK1) has been reported to induce fibrotic signaling in the setting of oxidative stress. However, the role of ASK1 and its mechanism of action in angiotensin II- (Ang II-) induced liver fibrosis remain largely unknown. METHODS: Human hepatic LX-2 stellate cells were treated with Ang II alone or cotreated with Ang II plus an ASK1 inhibitor (GS-4997) or siRNA-targeting ASK1. Immunofluorescent staining, real-time PCR, and western blotting were used to determine the expressionof α-SMA, Col I, and Col III expression. Cell viability was assessed by the CCK-8 assay. The concentrations of IL-1ß, IL-18, and TNF-α in conditioned medium were determined by ELISA. The levels of intracellular ROS in LX-2 cells were analyzed using a ROS assay kit. Exosome size was determined by electron microscopy. RESULTS: Ang II markedly increased the expression of extracellular matrix (ECM) proteins (α-SMA, Col I, and Col III) and proinflammatory cytokines (IL-1ß, IL-18, and TNF-α). Ang II also increased the expression of endoplasmic reticulum stress (ERS) markers (GRP78, p-PERK, and CHOP) and p-ASK1. Results also showed that pretreatment with GS-4997 or siRNA could abolish all the abovementioned effects on LX-2 cells. Furthermore, we found that exosome release caused by ASK1-mediated ERS was involved in the activation of LX-2 cells by Ang II. The activation of LX-2 cells could be blocked by treating the exosomes with annexin. CONCLUSIONS: In summary, we found that ASK1 mediates Ang II-activated ERS in HSCs and the subsequent activation of HSCs, suggesting a promising strategy for treating liver fibrosis.

Stable isotope labelling by amino acids in cell culture (SILAC) applied to quantitative proteomics of Edwardsiella tarda ATCC 15947 under prolonged cold stress.

Edwardsiella tarda poses a threat to human health and has resulted in enormous economic losses in aquaculture. Low temperatures are usually applied to contain the growth of this microorganism. In this study, stable isotope labelling by amino acids in cell culture (SILAC) was used to conduct comparative proteomic quantitation of E. tarda ATCC 15947 under cold stress for two weeks. We identified 1391 proteins, of which 898 were quantifiable. Of these, 72 proteins were upregulated and 164 were downregulated in response to cold stress. Even though E. tarda ATCC 15947 is not a psychrophile, several key proteins related to DNA synthesis and transcription were significantly upregulated. Additionally, proteins related to haemolytic activities and gluconeogenesis were upregulated, even though E. tarda ATCC 15497 is considered non-virulent in aquaculture. This study therefore delineated the specific proteomic response of this E. tarda ATCC 15947 to prolonged cold stress.

MiR-544 promotes immune escape through downregulation of NCR1/NKp46 via targeting RUNX3 in liver cancer.

OBJECTIVE: To study the potential role of miR-544 in the immune escape mechanism of hepatoma cells. METHODS: Natural killer (NK) cells were collected from healthy volunteers and patients with liver cancer. Interleukin (IL)-2 activated-NK-92 cells were transfected with miR-544 inhibitor/mimic or NC/pre-NC in HepG2 co-culture system. NK-92 cells were treated with control, IL-2, IL-2 + pre-NC, IL-2 + miR-544 mimic, IL-2 + miR-544 mimic + pcDNA and IL-2 + miR-544 mimic + pcDNA-runt-related transcription factor 3 (RUNX3) groups. Mice models of liver cancer were well established. Expression of miR-544, natural cytotoxicity receptor 1 (NCR1) and RUNX3 were evaluated by quantitative real-time PCR and western blotting. Flow cytometry and ELISA were used to determine NK cell cytotoxicity and the levels of INF-γ, respectively. RESULTS: MiR-544 was upregulated while NCR1 and RUNX3 was downregulated in NK cells of patients with liver cancer. The levels of IFN-γ and miR-544 expression were increased and decreased in IL-2 activated-NK cells, respectively. Inversely, miR-544 overexpression inhibited NK cell cytotoxicity by downregulating IFN-γ. However, miR-544 directly targeted RUNX3 and negatively regulated NCR1. Furthermore, miR-544 promoted immune escape of hepatoma cells in vivo and in vitro. CONCLUSION: miR-544 promoted the immune escape of liver cancer cells by downregulating NCR1 via targeting RUNX3.

Purification, characterization, and gene cloning of a new cold-adapted ß-galactosidase from Erwinia sp. E602 isolated in northeast China.

ß-Galactosidases are widely used in industry for elimination of lactose from milk products. A new ß-galactosidase was obtained from bacterial strain Erwinia sp. E602, newly isolated in northeast China. The enzyme was purified with the methods of ammonium sulfate fractionation, ion exchange, and gel filtration chromatography for further study of the enzymatic characteristics. The purified enzyme had a molecular weight of near 110 kDa. The optimum reaction temperature and pH of this enzyme was determined to be 40°C and 7.0, respectively, indicating that this enzyme was a mesophilic neutral ß-galactosidase. Furthermore, the enzyme retained near 10% of the activity at 0°C, which also suggested its cold-adapted property. Kinetics of the ß-galactosidase was studied, and the Km (Michaelis constant) and Vmax (maximum enzymatic reaction rate) of this enzyme were 0.21 mmol/L and 263.16 µmol/mg per minute, respectively. The effects of metal ions on the enzymatic activity and the lactose hydrolysis efficiency in milk, as well as its trans-glycosylation activity, were studied in this work. The ß-galactosidase coding gene was cloned to be a 3-kb length fragment, which shared at most 81% of identity with the published sequences in NCBI Blast database (https://blast.ncbi.nlm.nih.gov). Results in this work suggested it is a new ß-galactosidase and it has potential to be used in dairy and food processing.

Socioeconomic Burden of Pyogenic Liver Abscess Caused by Klebsiella Pneumoniae from a Teaching Hospital in East China.

Background: The prevalence of pyogenic liver abscess (PLA) is increasing worldwide. However, evaluation on its economic burden is still lack. Methods: A retrospective study that included all patients identified PLA from 2017 to 2020 was conducted. Clinical information and hospital costs were collected through the electronic medical records. We evaluated the economic burden using disability-adjusted life years (DALYs). Differences in socioeconomic burdens between Klebsiella pneumoniae-caused liver abscesses (KPLA) and non-Klebsiella pneumoniae-caused liver abscesses (non-KPLA) were compared. Results: We found 327 patients identified PLA in the study, including 146 with KPLA and 181 with non-KPLA. The demographic characteristics, median hospital stay, severity, and in-hospital mortality were similar between the two groups. The median total in-hospital cost was higher in the non-KPLA than in the KPLA group, although no statistical difference was found ($3607.2 vs $3424.6; P = 0.446). The median DALY loss was significantly higher in the KPLA than in the non-KPLA group [1.49 (0.97-2.30) vs 1.27 (0.87-1.89); P = 0.033)], and male patients presented a higher average DALY loss than female patients. KPLA had a substantially greater median indirect economic loss than the non-KPLA group [$1442.8 (915.9-17,221.5) vs $1232.5 (764.6-15,473.0); P = 0.028], and indirect economic loss exhibited a significant increase from 2017 to 2020 in patients with PLA. No differences were found in the socioeconomic burden between the two groups [$8019.6 (4200.3-21,832.1) vs $7436.4 (4023.2-19,063.9); P = 0.172]. Conclusion: The economic burden of PLA is significant, particularly in patients with KP. Patients with KPLA experienced increased DALY loss and indirect economic loss than non-KPLA. PLA must be prioritized as the indirect economic burden rises annually.

Olive oil authentication based on quantitative ß-carotene Raman spectra detection.

ß-carotene and oleic acids are important molecules to distinguish between extra olive oil (EVOO) and other oils. To identify adulteration which use common vegetable oils blended with ß-carotene to imitate EVOO, a non-invasive, lossless method is proved to be effective. The present work presents a laser confocal Raman technique for analyzing and comparing the differences of molecule between EVOO and SSO, which based on theoretical Raman spectra of ß-carotene, oleic acids and linoleic acids calculated by density functional theory (DFT). Chemometrics based on support vector regression (SVR) was used to realize quantitative analysis of ß-carotene in synthetic olive oils. Nine different volume ratios were prepared independently, and test set evaluation index of linear kernel of SVR as follow: RMSE 0.0653, R2 0.9868. The results show that laser confocal Raman technique, combined with theoretical Raman spectra based on DFT, could analyze composition of vegetable oil accurately, and identify low-cost imitation of olive oil.

Study on the Raman spectral characteristics of dynamic and static blood and its application in species identification.

This paper proposes a method to identify the blood of 4 poultry species (chicken, duck, goose and pigeon) based on Raman spectroscopy and its baseline. Samples were prepared by pretreatment methods of freezing, thawing, and dilution. The Raman spectra of dynamic blood and static blood were measured, respectively, and the spectral differences between the two research schemes were analyzed. The four species of poultry blood were identified based on the Raman spectroscopy and its baseline. The results show that the method can realize the identification of four species of poultry blood. In addition, the potential of Raman spectroscopy as a technique for determining carotenoids in blood has been clearly confirmed, which opens up the possibility to quickly determine whether poultry eats feed containing carotenoids without sample preparation.

Rapid detection of Chinese-specific peony seed oil by using confocal Raman spectroscopy and chemometrics.

Peony seed oil (PSO) is a new woody nut oil which is unique to China. Its unsaturated fatty acids are over 90% and are rich in α - linolenic acid. Although the PSO industry is in its infancy, it is bound to become a top vegetable oil food material because of its own advantages. The potential high commercial profit of its adulteration with cheap vegetable oil will be an important factor hindering the healthy development of PSO industry. It is of great significance to study the adulteration of PSO for preventing large-scale adulteration. In this study, the qualitative and quantitative analysis of PSO was realised based on Raman spectroscopy combined with chemometrics analysis, and the fatty acid composition of PSO was analysed according to Raman characteristic peaks. The technology can be applied to routine analysis and quality control of PSO.

Dynamic confocal Raman spectroscopy of flowing blood in bionic blood vessel.

How to quickly and safely identify blood species has always been an urgent problem for scientists. Smear test method has the risk of blood contamination, and the blood itself may carry some unknown viruses or pathogens, which will bring health risks to the testing personnel. Therefore, in order to meet the urgent needs of rapid and safe detection of blood, a technology which can detect dynamic confocal Raman spectroscopy of flowing blood in bionic blood vessel was proposed. The blood, which was sealed in the bionic blood vessel, flowed through the focus gaze area of laser by the microfluidic pump, to detect the dynamic blood Raman spectrum. Human blood and cattle blood were selected as experimental objects, and the experiments were carried out under the same parameters. Combined with PCA-LDA (principal component analysis and linear discriminate analysis) classification model, the predictive classification of the two species without error recognition was realized. The hidden weak Raman signals were mined by derivative spectra, and the fundamental differences of Raman spectra of two species were compared. Then the biochemical information that caused the differences was also analyzed. The results show the method can meet the detection requirements of sealed blood, and the Raman spectra of flowing blood is more representative than those of static blood.

Mangiferin alleviates endoplasmic reticulum stress in acute liver injury by regulating the miR-20a/miR-101a-Nrf2 axis.

This study aimed to investigate the mechanism of mangiferin on regulating endoplasmic reticulum (ER) stress in acute liver injury. The mouse model of acute liver injury was established by injection of LPS/D-GalN. The primary mouse hepatocytes were stimulated with LPS to induce the in vitro model. The effect of miR-20a/101a on the luciferase activity of Nrf2 3'-UTR was assessed by luciferase reporter assay. Mangiferin improved the liver function, inhibited the oxidative stress and ER stress and down-regulated the expressions of miR-20a and miR-101a in LPS/D-GalN-induced mice and LPS-induced hepatocytes. The knockdown of miR-20a and miR-101a co-operatively alleviated ER stress of LPS-induced hepatocytes. miR-20a and miR-101a both targeted Nrf2 and the over-expression of miR-20a or miR-101a decreased Nrf2 protein level, while their silences increased Nrf2 protein level. The silence of miR-20a and miR-101a promoted Nrf2 expression and inhibited the ER stress in LPS-induced hepatocytes, while the knockdown of Nrf2 reversed these effects. The over-expression of miR-20a and miR-101a eliminated the effects of mangiferin on Nrf2 protein level and ER stress in LPS-induced hepatocytes and Nrf2 over-expression altered these trends. Our findings suggest that mangiferin alleviates ER stress in acute liver injury by regulating the miR-20a/miR-101a-Nrf2 axis.

Analyzing hedyotis diffusa mechanisms of action from the genomics perspective.

BACKGROUND AND OBJECTIVE: Hedyotis diffusa is an herb used for anti-cancer, anti-oxidant, anti-inflammatory, and anti-fibroblast treatment in the clinical practice of Traditional Chinese Medicine. However, its pharmacological mechanisms have not been fully established and there is a lack of modern scientific verification. One of the best ways to further understand Hedyotis diffusa's mechanisms of action is to analyze it from the genomics perspective. METHODS: In this study, we used network pharmacology approaches to infer the herb-gene interactions, the herb-pathway interactions, and the gene families. We then analyzed Hedyotis diffusa's mechanisms of action using the genomics context combined with the Traditional Chinese Medicine clinical practice and the pharmacological research. RESULTS: The results obtained in the pathway and gene family analysis were consistent with the Traditional Chinese Medicine clinical experience and the pharmacological activities of Hedyotis diffusa. CONCLUSIONS: Our approach can identify related genes and pathways correctly with little a priori knowledge, and provide potential directions to facilitate further research.

LncRNA GAS5 enhanced the killing effect of NK cell on liver cancer through regulating miR-544/RUNX3.

This study aimed to explore the role of lncRNA GAS5 in the regulation of the killing effect of NK cells on liver cancer. Compared with a control group, lncRNA GAS5, RUNX3, and NCR1 were down-regulated in NK cells of patients with liver cancer, whereas miR-544 expression was up-regulated in NK cells of patients with liver cancer. Activated NK cells had higher IFN-γ level. Knockdown of GAS5 in activated NK cells decreased IFN-γ secretion, NK cell cytotoxicity, the percentage of CD107a+ NK cells, and the apoptosis rate of HepG2 and Huh7 cells. We also proved the interaction of GAS5 and miR-544, and the negative regulation role of GAS5 on miR-544. GAS5 overexpression in activated NK cells increased RUNX3 expression, IFN-γ secretion, the NK cell cytotoxicity, the percentage of CD107a+ NK cells, and the apoptosis rate of HepG2 cells, while miR-544 mimic abolished the promotion effect of GAS5 overexpression. Finally, in vivo experiments indicated an inhibition effect of GAS5 in tumor growth. LncRNA GAS5 overexpression enhances the killing effect of NK cell on liver cancer through regulating miR-544/RUNX3.

[Mechanism of liver injury in severe acute pancreatitis rats and role of melatonin].

OBJECTIVE: To investigate the activation level of nuclear factor-kappaB (NF-kappaB) in liver injury caused by severe acute pancreatitis (SAP) and the protective role of melatonin against liver injury. METHODS: Ninety-six Sprague-Dawley (SD) rats were randomly divided into 3 equal groups: SAP group undergoing injection of sodium taurocholate to establish SAP models, melatonin (Mel) treatment group undergoing intraperitoneal injection 50 mg/kg 30 minutes before the establishment of SAP models, and sham operation group (Sham group). 4, 12, 24, and 48 hours after the onset of operation, blood samples were collected from the inferior vena casa of 8 rats from each group to measure the serum level of amylase (AMY) and alanine transaminase (ALT) by iodine colorimetry, and to detect the serum level of tumor necrosis factor-alpha (TNF-alpha) by ELISA. The livers were taken out to undergo pathological examination. Immunohistochemistry was used to examine the percentage of nuclear factor (NF)-kappaB in the hepatocytes. Terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) was used to determine the extent of hepatic apoptosis. RESULTS: The AMY and ALT levels at different time points of the SAP and Mel subgroups were all significantly higher than those of the sham operation subgroups (all P<0.05), and the AMY and ALT levels at different time points of the Mel subgroup were all significantly lower than those of the SAP subgroups (all P<0.05). The liver NF-kappaB activation level and hepatocellular apoptosis index of the SAP group increased since the fourth hour after the operation, and peaked at the time point of 24 hour, all significantly higher than those of the sham operation group (all P<0.05), and then declined. The TNF-alpha level at the time points of 12, 24, and 48 h in the SAP group were all significantly higher than those of the other 2 groups (all P<0.05). The levels of TNF-alpha, AMY, and ALT, the activity of NF-kappaB, and the extent of hepatocellular apoptosis at any time points of the Mel group were all significantly lower than those of the SAP group, but significantly higher than those of the sham operation group. Microscopy showed that the liver pathological damages of the Mel group were milder than those of the SAP group. CONCLUSION: SAP with liver injury is associated with the hepatic NF-kappaB activation leading to the production of NF-kappaB dependent cytokines and chemokines such as TNF-alpha. Melatonin reduces the apoptosis and necrosis in liver by inhibiting the activity of NF-kappaB and decreasing the expression of TNF-alpha.

[Knockdown of STAT3 inhibits proliferation and migration of HepG2 hepatoma cells induced by IFN1].

Objective To prepare lentiviruses expressing shRNA sequences targeting human signal transducer and activator of transcription 3 (STAT3) and detect the effect of STAT3 knockdown on type I interferon (IFN1)-induced proliferation and migration in HepG2 cells. Methods Four STAT3-targeting shRNA sequences (shRNA1-shRNA4) and one control sequence (Ctrl shRNA) were selected and cloned respectively into pLKO.1-sp6-pgk-GFP to construct shRNA-expressing vectors. Along with backbone psPAX2 and pMD2.G vectors, they were separately transfected into HEK293T cells to prepare lentiviruses. HepG2 cells were infected with the lentiviruses. Cytoplastic STAT3 level was detected by Western blotting to screen effective shRNA sequence(s) targeting STAT3. Proliferation and migration of HepG2 cells were analyzed by CCK-8 assay and TranswellTM migration and scratching assay, respectively. To detect the effect of IFN1 on cell proliferation and migration of HepG2 cells, the cells were treated with 2000 U/mL IFNα2b for indicated time and the activation of IFN-triggered STAT1 signal transduction was assayed by Western blotting. Results Two most effective STAT3-targeting shRNA sequences shRNA1 and shRNA2 were selected, and the expression of both STAT3 shRNA significantly decreased proliferation and migration of HepG2 cells. When treated with IFNα2b, 2000 U/mL of IFN1 showed more competent in attenuating growth and migration of HepG2 cells. Our data further proved that knockdown of STAT3 increased the phosphorylation of STAT1, and IFNα2b further enhanced the activation of STAT1 signaling in HepG2 cells. Conclusion Knockdown of STAT3 inhibits cell migration and growth, and rescues IFN response through up-regulating STAT1 signal transduction in HepG2 hepatoma cells.